Reaction of Bacillus subtilis glutamine phosphoribosylpyrophosphate amidotransferase with oxygen: chemistry and regulation by ligands.
The inactivation of glutamine phosphoribosylpyrophosphate amidotransferase by reaction of its iron-sulfur center with O2 is believed to be a physiologically important mode of regulation of this enzyme in Bacillus subtilis cells in the stationary phase of growth. Chemical and physical changes accompanying oxidation of the purified enzyme by O2 were studied. The iron of the 4Fe-4S center was oxidized to enzyme-bound high-spin Fe3+; the S2- was oxidized to a mixture of S0 bound as thiocystine and unidentified products. The oxidant appeared to be O2, rather than peroxide, superoxide, hydroxyl radical, or singlet oxygen. Gross physical changes in the oxidized enzyme were shown by its aggregation, decreased solubility, and altered circular dichroic spectrum. Experimental variables affecting the rate of oxidative inactivation were described; the most important of these was modulation of rates of inactivation by the allosteric inhibitors AMP, ADP, GMP, GDP and by the substrate P-Rib-PP. AMP was a potent stabilizer, whose effect was antagonized by P-Rib-PP. The other nucleotides, either acting singly or acting as synergistic pairs, were destabilizers and able to antagonize stabilization by AMP. The results are discussed in terms of the regulation of the stability of amidotransferase and its degradation in vivo.
D000227:Adenine Nucleotides / Q000378:metabolism; D000582:Amidophosphoribosyltransferase / Q000037:antagonists & inhibitors*; D001412:Bacillus subtilis / Q000201:enzymology*; D002942:Circular Dichroism; D006150:Guanine Nucleotides / Q000378:metabolism; D007506:Iron-Sulfur Proteins / Q000378:metabolism; D008956:Models, Chemical; D010100:Oxygen / Q000494:pharmacology*; D010430:Pentosyltransferases / Q000037:antagonists & inhibitors*
D000227:Adenine Nucleotides; D006150:Guanine Nucleotides; D007506:Iron-Sulfur Proteins; D010430:Pentosyltransferases; D000582:Amidophosphoribosyltransferase; D010100:Oxygen